An inactive Zika NS2B-NS3pro protease construct for investigating allosteric inhibitors

Area of Science:

• Virology
• Structural Biology
• Drug Discovery

Background:

• The Zika virus protease (NS2B-NS3) is crucial for viral maturation and a key therapeutic target.
• Developing direct active site inhibitors is challenging due to the protease's hydrophilic nature.
• Allosteric inhibition presents a promising alternative strategy.

Purpose of the Study:

• To engineer a stable Zika virus protease construct suitable for allosteric inhibitor screening.
• To identify novel allosteric binding sites and fragments for therapeutic development.

Main Methods:

• Engineered an NS2B-NS3 protease construct with a glycine-rich linker.
• Determined the crystal structure and solution NMR spectrum of the construct.
• Performed fragment screening to identify potential allosteric inhibitors.

Main Results:

• Successfully created and characterized a novel NS2B-NS3 protease construct.
• Identified two new fragments that bind to the engineered protease construct.
• The construct design facilitates the study of allosteric inhibition by excluding a conformationally flexible region.

Conclusions:

• The engineered NS2B-NS3 protease construct is a valuable tool for screening and characterizing allosteric inhibitors.
• This strategy advances the development of therapeutics targeting Zika virus protease.
• Identified fragments serve as starting points for developing novel antiviral agents.